3-Methylene flavanones and 3-methylene chromanones

ABSTRACT

Compounds which are 3-methylene flavanones and 3-methylene chromanones having activity against microorganisms are disclosed. The compounds are represented by the general structural formula: ##STR1## wherein: R 1  is a member selected from the group consisting of hydrogen, Br, CH 3  and OCH 3  ; R 2  is selected from the group consisting of hydrogen and ##STR2## wherein R 4  is a member selected from the group consisting of hydrogen, Br, Cl, CH 3 , OCH 3 , NO 2 , N(CH 3 ) 3  and CN; R 5  is selected from the group consisting of hydrogen and Cl, with the proviso that when R 5  is Cl, R 4  is hydrogen or Cl; and R 3  is selected from the group consisting of hydrogen, phenyl, 2-thienyl, 4-pyridyl and naphthyl, with the proviso that when R 3  is naphthyl, R 1  and R 2  are hydrogen.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to 3-methylene flavanones and 3-methylenechromanones which exhibit activity against bacteria, yeast or fungi.

2. Prior Art

The prior art discloses numerous derivatives of flavanone (flavone).Flavanone has the following structural formula and is numbered as shown:##STR3## Chromanone has the following structure: ##STR4## U.S. Pat. No.3,002,797 discloses flavanone-7-oxy-acetamides. U.S. Pat. No. 3,495,009discloses antiinflammatory flavanone compounds, unsubstituted at the3-position, and halogenated in the 4' position. U.S. Pat. No. 3,598,840discloses the production of flavanones having a hydroxy or acetoxyradical in the 3-position. U.S. Pat. Nos. 3,410,851 and 3,753,985 bothassigned to the present assignee, disclose 3-aminomethyl substitutedflavanones which are antimicrobial agents.

None of the prior art referred to above discloses flavanone orchromanone antimicrobial compounds having a methylene group at the3-position.

SUMMARY OF THE INVENTION

The present invention is directed to 3-methylene flavanones and3-methylene chromanones The claimed compounds can be represented bygeneral structural formula I as: ##STR5## and pharmacologicallyacceptable, non-toxic salts thereof wherein: R¹ is a member selectedfrom the group consisting of hydrogen, Br, CH₃ and OCH₃ ; R² is selectedfrom the group consisting of hydrogen and ##STR6## wherein R⁴ is amember selected from the group consisting of hydrogen, Br, Cl, CH₃,OCH₃, NO₂, N(CH₃)₂, C(CH₃)₃ and CN; R⁵ is selected from the groupconsisting of hydrogen and Cl, with the proviso that when R⁵ is Cl, R⁴is hydrogen or Cl; and R³ is selected from the group consisting ofhydrogen, phenyl, 2-thienyl, 4-pyridyl and naphthyl, with the provisothat when R³ is naphthyl, R¹ and R² are hydrogen.

DESCRIPTION OF THE INVENTION

In general, the compounds of the present invention can be convenientlyplaced in the following three categories:

A. 3-Methylene Flavanones, where R³ is hydrogen; ##STR7## B. 3-MethyleneFlavanones, where R¹, R⁴ and R⁵ are hydrogen and R³ is phenyl, 2-thienylor 4-pyridyl; ##STR8## C. 3-Methylene Chromanones, where R² is hydrogenand R³ is naphthyl; ##STR9##

Two methods were used to prepare the 3-methylene flavanones andchromanones. In the first method, the appropriately-substituted2-hydroxyacetophenones V are condensed with substituted benzaldehydes VIor naphthaldehydes by heating in aqueous ethanol in the presence of abase such as borax or sodium hydroxide to give the correspondingchalcones VII. The 2-hydroxyacetophenones are the products of the Friesrearrangement ("Preparative Organic Chemistry", G. Hilgetag and A.Martini Eds., John Wiley & Sons, New York, New York, 1972, page 1068)and are readily available. In some cases (e.g., Example 14) thechalcones VII spontaneously cyclize to the flavanones VIII. In othercases (e.g., Example 4) the chalcones VII were cyclized to theflavanones VIII by heating them in dilute acid. The flavanones andchromanones can be converted to the corresponding Mannich bases IX bytreatment with formaldehyde, dimethylamine and hydrochloric acid (See B.Reichert "Die Mannich Reaktion", Berlin, 1959, p. 22-24). These Mannichbases decompose on treatment with dilute base (e.g., Example 4) to yieldthe desired 3-methylene flavanones II and III or desired 3-methylenechromones IV.

Alternately, the flavanones VIII can be converted directly to the3-methylene flavanones by heating for several hours with the reagentbis-(dimethylamino)methane in acetic anhydride. ##STR10##

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following compounds comprise preferred embodiments of Formula III:##STR11##

                  TABLE A                                                         ______________________________________                                        3-Methylene Flavanones where R.sup.3 is hydrogen                              Example No.                                                                             TR Number   R.sup.1  R.sup.4  R.sup.5                               ______________________________________                                        1         3489        H        H        H                                     2         3667        H        3'-Cl    H                                     3         3498        H        H        4'-Cl                                 4         3734        H        2'-Cl    4'-Cl                                 5         3671        H        3'-Br    H                                     6         3673        H        4'-Br    H                                     7         3648        H        3'-OCH.sub.3                                                                           H                                     8         3762        H        4'-OCH.sub.3                                                                           H                                     9         3666        H        3'-NO.sub.2                                                                            H                                     10        3614        H        4'-NO.sub.2                                                                            H                                     11        3654        H        4'-N(CH.sub.3).sub.2                                                                   H                                     12        3670        H        3'-CH.sub.3                                                                            H                                     13        3485        H        4'-CH.sub.3                                                                            H                                     14        3683        H        4'-C(CH.sub.3).sub.3                                                                   H                                     15        3665        H        3'-CN    H                                     16        3649        H        4'-CN    H                                     17        3791        6-Br     4'-Br    H                                     18        3782        7-OCH.sub.3                                                                            4'-Cl    H                                     19        3783        6-CH.sub.3                                                                             H        H                                     ______________________________________                                    

EXAMPLE 1 3-methylene flavanone

A mixture of 44.8 g (0.2 mol) of flavanone (commercially available fromAldrich Chemical Co., Milwaukee, Wisconsin), 32.6 g (0.4 mole) ofdimethylamine hydrochloride, 12.6 g (0.4 mol) of paraformaldehyde, 1 mlof concentrated HCl, and 120 ml of 2-propanol was stirred at reflux for1 hour. The reaction was then allowed to cool to room temperature, thesolvent was removed under reduced pressure and the residue partitionedbetween 400 ml of ether and 1 liter of 5 percent aqueous hydrochloricacid. The aqueous phase was separated, washed with 200 ml of ether,cooled and made basic with solid K₂ CO₃. The liberated base wasextracted into ether, the ether solution dried over anhydrous Na₂ SO₄,filtered and concentrated under reduced pressure. The residue was takenup in 200 ml of ethyl acetate and treated with excess acetyl chlorideand ethanol. Upon cooling, white crystals precipitated. After drying, ayield of 43.8 g of 3-dimethylaminomethyl flavanone hydrochloride, (mp172°-174° C.) was obtained.

Ten grams of this hydrochloride salt was dissolved in 400 ml of H₂ O andheated on the steam bath for 2 hours. After cooling to room temperature,the cloudy solution was extracted with ether. The ether solution wasdried over anhydrous Na₂ SO₄, filtered and evaporated under reducedpressure to give 2.2 g of an oil. The oil was taken up in hot petroleumether (b.p. 30° C.), filtered, and concentrated to a 10 ml volume. Aftercooling to room temperature, the solution was decanted from a smallamount of gummy precipitate. Evaporation of the solution gave 300 mg ofa yellow oil of 3-methylene flavanone.

NMR Spectrum (CCl₄): δ 5.05 (1H, t, J=2 Hz); 5.85 (1H, t, J=2 Hz); 6.3(1H, t, J=2 Hz).

Mass Spectrum (70 eV) m/e: 236 (M+).

Infrared Spectrum (CCl₄): 1670 cm⁻¹ (νco).

EXAMPLE 2 3'-chloro-3-methylene flavanone

3'-Chloro flavanone was prepared as described in J. Taiwan Pharm. Assoc.3, 39 (1951) and Chem Abst. 49, 2432 g (1955). A 18.5 g (0.072 mol)portion of 3'-chloro flavanone was converted to 9.9 g of3'-chloro-3-dimethylaminomethyl flavanone hydrochloride (m.p. 109°-151°C.) in a similar manner as described in Example 1.

Calcd for C₁₈ H₁₈ ClNO₂.HCl: C, 61.37; H, 5.44; N, 3.98. Found: C,61.19; H, 5.37; N, 3.86.

A mixture of 1.76 g (5 mmol) of this hydrochloride salt and 12 ml ofbis-(dimethylamino)methane (commercially available from Aldrich ChemicalCo.) was stirred under argon at room temperature; 8 ml of aceticanhydride was added dropwise. A transient rise in temperature occurred,which was stopped by immersing the reaction vessel in a cooling bath.After 20 minutes, an additional 4 ml portion of acetic anhydride wasadded. The reaction mixture was then poured into 100 g of crushed iceand extracted with ether. The ether phase was separated, washed withsodium bicarbonate solution, dried over anhydrous Na₂ SO₄, filtered andevaporated to give 1.5 g of a yellow oil. The oil was chromatographed on100 g of silica gel and eluted with benzene to yield 1 g of a paleyellow waxy solid of 3'-chloro-3-methylene flavanone, m.p. 85°-88° C.

NMR spectrum (CDCl₃): δ 5.2 (1H, m); 6.0 (1H, m); 6.5 (1H, m). Massspectrum (70 eV) m/e: Calcd for C₁₆ H₁₁ ClO₂ : 270.04469. Found:270.0421 (M+).

Infrared Spectrum (CHCl₃): 1675 cm⁻¹ (νco).

EXAMPLE 3 4'-chloro-3-methylene flavanone

In a similar manner as described in Example 1, 10 g of 4'-chloroflavanone, as described in Bull. Soc. Chim. France 2248 (1963), wasconverted to 4.0 g of crude 4'-chloro-3-dimethylaminomethyl flavanonehydrochloride. The hydrochloride salt was recrystallized twice fromacetone to give 1.1 g of crystals, m.p. 147°-149° C.

Calcd. for C₁₈ H₁₈ ClNO₂.HCl: C, 61.37; H, 5.44; N, 3.98. Found: C,61.88; H, 5.58; N, 4.12.

The combined filtrates from the above recrystallizations were mixed withH₂ O and stirred for one hour. An oil separated that later crystallized.The crystals were filtered and recrystallized from petroleum ether togive 500 mg of the desired flavanone as a white solid, m.p. 84°-85° C.

Calcd for C₁₆ H₁₁ ClO₂ : C, 70.98; H, 4.10. Found: C, 70.56; H, 4.23.

NMR spectrum (CCl₄): δ 5.1 (m, 1H), 5.90 (m, 1H), 6.3 (m, 1H).

EXAMPLE 4 2',4'-dichloro-3-methylene flavanone

2-Hydroxy-2',4'-dichlorochalcone, as described in Arch. Pharm. 295, 16(1962), was cyclized to the flavanone by the procedure described inBull. Soc. Chem. France 674 (1973). Two recrystallizations from methanolgave 3 g of 2',4'-dichloro flavanone as pale tan crystals, m.p. 91°-92°C.

Calcd. for: C₁₅ H₁₀ Cl₂ O₂ : C, 61.46; H, 3.44. Found: C, 61.42; H,3.46.

A mixture of 3 g (10 mmol) of the 2',4'-dichloro flavanone and 27 ml ofbis-(dimethylamino)methane was stirred at room temperature; 27 ml ofacetic anhydride was added dropwise over 15 minutes. A slighttemperature rise occurred. One hour later the reaction was partitionedbetween 200 ml of ether and 200 ml of cold H₂ O. The aqueous phase wasseparated, washed with two 250 ml portions of ether, and discarded. Thecombined ether extracts were washed with saturated aqueous NaHCO₃solution, dried over anhydrous Na₂ SO₄, filtered and evaporated. Theresidual oil was chromatographed on 200 g of silica gel, eluting withbenzene. This gave 2.5 g of 2',4'-dichloro-3-methylene flavanone,isolated as a solid, m.p. 86°-87° C.

Calcd. for C₁₆ H₁₀ Cl₂ O₂ : C, 62.97; H, 3.30. Found: C, 63.20; H, 3.36.

NMR Spectrum (CCl₄): δ 4.9 (1H, m), 6.3 (2H, m).

EXAMPLE 5 3'-bromo-3-methylene flavanone

In a similar manner as described in Example 1, 17 g of 3'-bromoflavanone, as described in J. Chinese Chem. Soc. 11, 102 (1964); ibid.10, 125 (1963) and Chem. Abst. 62, 6011b (1965), was converted to 6.7 gof the hydrochloride salt of 3'-bromo-3-dimethylaminomethyl flavanone,as white crystals, m.p. 105°-172° C.

Calcd. for C₁₈ H₁₈ BrNO₂.HCl: C, 54.49; H, 4.83, N, 3.53. Found: C,54.27; H, 4.93; N, 3.79.

A 1.91 g portion of this hydrochloride salt was converted to 1.3 g of3'-bromo-3-methylene flavanone, m.p. 98°-100° C., using the proceduredescribed in Example 2.

Calcd. for C₁₆ H₁₁ BrO₂ : C, 60.97; H, 3.52. Found: C, 60.80; H, 3.56.

NMR Spectrum (CDCl₃): δ 5.3 (1H, m), 6.0 (1H, m), 6.5 (1H, m).

EXAMPLE 6 4'-bromo-3-methylene flavanone

In a similar manner as described in Example 1, 11.8 g of 4'-bromoflavanone, as described in J. Taiwan Pharm. Assoc. 3, 39 (1951) andChem. Abst. 49, 2432 g (1955), was converted to 4.2 g of thehydrochloride salt of 4'-bromo-3-dimethylaminomethyl flavanone, m.p.157°-159° C.

Calcd. for C₁₈ H₁₈ BrNO₂.HCl: C, 54.49; H, 4.83; N, 3.53. Found: C,54.42; H, 4.93; N, 3.63.

A 1.91 g portion of this hydrochloride salt was converted to 1.28 g of4'-bromo-3-methylene flavanone, m.p. 91°-92° C., using the proceduredescribed in Example 2.

Calcd. for C₁₆ H₁₁ BrO₂ : C, 60.97; H, 3.52. Found: C, 61.05; H, 3.42.

NMR Spectrum (CDCl₃): δ 5.1 (1H, m), 5.9 (1H, m), 6.4 (1H, m).

EXAMPLE 7 3'-methoxy-3-methylene flavanone

In a similar manner as described in Example 1, 25 g of 3'-methoxyflavanone, as described in Chem. Ber. 38, 933 (1905), was converted tothe hydrochloride salt of 3'-methoxy-3-dimethylaminomethyl flavanone,m.p. 155°-156° C., 3.65 g, recrystallized from ethyl acetate.

Calcd. for C₁₉ H₂₁ NO₃.HCl: C, 65.61; H, 6.38; N, 4.03. Found: C, 65.31;H, 6.62; N, 4.21.

The filtrate from the crystallization was evaporated to give 2 g of anoil. This was partitioned between ether and saturated aqueous NaHCO₃solution. The ether phase was separated, dried over anhydrous Na₂ SO₄,filtered, and evaporated. The oily residue was taken up in 10 ml ofmethanol and 5 ml of methyl iodide and allowed to stand for severaldays. The solvent was then removed and the gummy solid residue waspartitioned between ether and 10 percent NaOH solution. The ether phasewas separated, dried over anhydrous Na₂ SO₄, filtered and concentratedto give 700 mg of an oil. The oil was taken back up in ether and washedwith 5 percent HCl solution, H₂ O, then dried over anhydrous Na₂ SO₄,filtered and evaporated to give 480 mg of solid. The solid was taken upin benzene and purified by preparative thin-layer chromatography (tlc);the silica gel tlc plates were manufactured by E. Merck & Co.,Darmstadt, Germany. 200 Mg of the desired product, 3'-methoxy-3-methylene flavanone, was obtained.

NMR Spectrum (CCl₄): δ 5.1 (1H, m), 5.9 (1H, m), 6.3 (1H, m).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₇ H₁₄ O₃ : 266.0930. Found:266.0937 (M+).

Infrared Spectrum (CCl₄): 1680 cm⁻¹ (νco).

EXAMPLE 8 4'-methoxy-3-methylene flavanone

In a similar manner as described in Example 1, 27.6 g of 4'-methoxyflavanone prepared as described in Acta Phytochem. 2, 99 (1925) andChem. Abst. 20, 2162 (1926) was converted to 2.85 g of the hydrochloridesalt of 4'-methoxy-3-dimethylaminomethyl flavanone, m.p. 149°-151° C.

Calcd. for C₁₉ H₂₁ NO₃.HCl: C, 65.61; H, 6.38; N, 4.03. Found: C, 65.40;H, 6.46; N, 4.06.

2.0 g of the hydrochloride salt was converted to 1.44 g of4'-methoxy-3-methylene flavanone, isolated as a yellow oil, using theprocedure described in Example 2.

NMR Spectrum (CCl₄): δ 5.1 (1H, m), 5.9 (1H, m), 6.3 (1H, m).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₇ H₁₄ O₃ : 266.0939. Found:266.09161 (M+).

Infrared Spectrum (CCl₄): 1685 cm⁻¹ (νco).

EXAMPLE 9 3'-nitro-3-methylene flavanone

A mixture of 1.35 g (5 mmol) of 3'-nitroflavanone [prepared as describedin J. Chem. Soc. Japan, Pure Chem. Sect. 74, 827 (1953)], and 12 ml ofbis-(dimethylamino)methane was stirred at room temperature; 12 ml ofacetic anhydride was added dropwise over 15 minutes. The temperaturerose to 77°. After cooling to ambient temperature, the reaction waspartitioned between 100 ml portions of ether and 100 ml portions of coldwater. The aqueous phase was separated, washed with two 100 ml of ether,and discarded. The combined ether extracts were washed with saturatedaqueous NaHCO₃ solution, dried over anhydrous Na₂ SO₄, filtered andevaporated. The residual oil was chromatographed on 100 g of silica gel,eluting with benzene to yield 548 mg of 3'-nitro-3-methylene flavanone,isolated as a white solid, m.p. 131°-132° C.

NMR Spectrum (CDCl₃): δ 5.3 (1H, m), 6.2 (1H, m), 6.6 (1H, m).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₆ H₁₁ NO₄ : 281.06873. Found:281.068700 (M+).

Infrared Spectrum (CDCl₃): 1680 cm⁻¹ (νco).

EXAMPLE 10 4'-nitro-3-methylene flavanone

A mixture of 18.1 g (67 mmol) of 4'-nitro flavanone, prepared asdescribed in J. Chem. Soc. Japan, Pure Chem. Sect. 74, 827 (1953), 4.04g (134 mmol) of paraformaldehyde, 10.97 g (134 mmol) of dimethylaminehydrochloride, 1 ml of concentrated HCl, and 200 ml of 2-propanol wasrefluxed for three hours. When cool, the solvent was removed underreduced pressure and the residue partitioned between CHCl₃ and 5 percentaqueous HCl. The layers were separated and the organic layer was washedwith 3-100 ml portions of 5 percent HCl. The combined aqueous extractswere neutralized with solid Na₂ CO₃ and extracted with ether. The etherwas dried over anhydrous Na₂ SO₄, filtered, and concentrated. The oilyresidue was dissolved in ethyl acetate and treated with 1 ml of ethanoland 1.5 ml of acetyl chloride. The solvent was then evaporated to give aglassy residue. This was taken up in 500 ml of hot ethyl acetatecontaining 20 ml of acetone and cooled. A precipitate formed that wasrecrystallized from methanol to give 810 mg of 4'-nitro-3-methyleneflavanone as a solid, m.p. 135°-136° C.

Calcd. for C₁₆ H₁₁ NO₄ : C. 68.31; H, 3.94; N, 4.98. Found: C, 67.68; H,3.98; N, 5.09.

NMR Spectrum (CDCl₃): δ 5.3 (s, 1H), 6.2 (s, 1H), 6.5 (s, 1H). EXAMPLE11

4'-dimethylamino-3-methylene flavanone

In a similar manner as described in Example 1, 16.5 g of4'-dimethylamino flavanone, prepared as described in J. Chem. Soc.Japan, Pure Chem. Sect. 74, 827 (1953), was converted to 2.6 g of thehydrochloride salt of 4'-dimethylamino-3-dimethylaminomethyl flavanone,recrystallized from alcohol, m.p. 184°-186° C.

Calcd. for C₂₀ H₂₄ N₂ O₂.HCl: C, 66.56; H, 6.98; N, 7.76. Found: C,66.27; H, 7.11; N, 7.53.

The filtrates from the recrystallization of the HCl salt were evaporatedunder reduced pressure. The oily residue was partitioned between 200 mleach of H₂ O and ether. The ether phase was separated, dried overanhydrous Na₂ SO₄, filtered and evaporated. This residue was extractedtwice with 250 ml portions of boiling pentane, and the combined extractscooled in the refrigerator. A yellow precipitate formed that wasseparated and recrystallized from CHCl₃ -petroleum ether to give 1.5 gof 4'-dimethylamino-3-methylene flavanone as a solid, m.p. 110°-112° C.

Calcd for C₁₈ H₁₇ NO₂ : C, 77.39; H, 6.13; N, 5.01. Found: C, 76.16; H,7.25; N, 5.25.

NMR Spectrum (CDCl₃): δ 2.9 (s, 6H), 5.3 (d, 1H, J=4 Hz), 5.4 (d, 1H,J=4 Hz).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₈ H₁₇ NO₂ : 279.1255. Found:279.1269 (M+).

EXAMPLE 12 3'-methyl-3-methylene flavanone

A mixture of 3-methylbenzaldehyde (12 g, 0.1 mol), 2-hydroxyacetophenone(13.6 g, 0.1 mol), and 2 liters of 0.1 N NaOH was stirred at roomtemperature for 4 days. The reaction was extracted with ether and theether dried and evaporated to give 20.5 g of 3'-methyl flavanone as anoil. The oil, not further purified but used directly, was converted in asimilar manner as described in Example 1 into 8.2 g of the hydrochloridesalt of 3'-methyl-3-dimethylaminomethyl flavanone, as a solid, m.p.156°-158° C.

Calcd. for C₁₉ H₂₁ NO₂.HCl: C. 68.77; H, 6.68; N, 4.22. Found: C. 67.84;H, 6.31; N, 4.22.

1.66 g of this hydrochloride salt was converted to 230 mg of3'-methyl-3-methylene flavanone, isolated as an oil, using the proceduredescribed in Example 2.

NMR Spectrum (CDCl₃): δ 2.4 (3H, s), 5.2 (1H, m), 6.0 (1H, m), 6.4 (1H,m).

Mass Spectrum (70 eV) m/e: Calcd: for C₁₇ H₁₄ O₂ : 250.09930. Found:250.09953 (M+).

Infrared Spectrum (CHCl₃): 1680 cm⁻¹ (νco).

EXAMPLE 13 4'-methyl-3-methylene flavanone

A mixture of 12 g (0.1 mol) of 4-methylbenzaldehyde, 13.6 g (0.1 mol) of2-hydroxyacetophenone, 250 ml of ethanol, and 250 ml of H₂ O containing38.1 g (0.1 mol) of sodium borate was refluxed overnight. It was cooled,concentrated under reduced pressure, and the residue extracted withCHCl₃. The CHCl₃ was dried over anhydrous Na₂ SO₄, filtered, andevaporated to give a mixture from which was extracted 13.6 g of impure4'-methyl flavanone [See German Offen. No. 2,535,338 (17 Feb. 1977)].The material was not further purified but was converted directly, in asimilar manner as described in Example 1, to the HCl salt of4'-methyl-3-dimethylaminomethyl flavanone, as a solid recrystallizedfrom methanol-ethyl acetate, m.p. 167°-169° C.

Calcd. for C₁₉ H₁₂ NO₂.HCl: C, 68.77; H, 6.68; N, 4.22. Found: C, 68.74;H, 6.76; N, 4.28.

The filtrates from the recrystallization of the HCl salt were evaporatedunder reduced pressure and the residue treated with boiling ethylacetate. The ethyl acetate solution was cooled, filtered, and evaporatedto give an orange oil. It was treated in the manner of Example 7 to give450 mg of 4'-methyl-3-methylene flavanone isolated as an oil.

Calcd. for C₁₇ H₁₄ O₂ : C, 81.58; H, 6.64. Found: C, 81.09; H, 5.68.

NMR Spectrum (CCl₄): δ 2.4 (s, 3H), 5.2 (m, 1H), 5.9 (m, 1H), 6.4 (m,1H).

EXAMPLE 14 4'-t-butyl-3-methylene flavanone

A mixture of 22.7 g (0.14 mol) of 4-t-butylbenzaldehyde, 19 g (0.14 mol)of 2-hydroxyacetophenone, 38 g (0.1 mol) of borax, 250 ml of H₂ O and150 ml of ethanol was refluxed for one day. The reaction was cooled anddiluted with an equal volume of H₂ O and extracted with ether. The etherwas washed with brine, dried over anhydrous Na₂ SO₄, filtered, andevaporated to give a brown oil. The oil was chromatographed on 1200 g ofsilica gel, eluting with benzene. The desired product was obtained as anoil. Recrystallization from petroleum ether gave 2.8 g of 4'-t-butylflavanone as white crystals, m.p. 103°-104° C.

Calcd. for C₁₉ H₂₀ O₂ : C, 81.39; H, 7.19. Found: C, 81.26; H, 6.88.

In a similar manner as described in Example 4, 1.40 g of this flavanonewas converted to 112 mg of 4'-t-butyl-3-methylene flavanone, isolated asa pale yellow solid, m.p. 104°-109° C.

NMR Spectrum (CDCl₃): δ 1.3 (9H, s), 5.2 (1H, m), 5.9 (1H, m), 6.4 (1H,m).

Mass Spectrum (70 eV) m/e: Calcd. for C₂₀ H₂₀ O₂ : 292.14622. Found:292.15531 (M+).

Infrared Spectrum (CHCl₃): 1680 cm⁻¹ (νco).

EXAMPLE 15 3'-cyano-3-methylene flavanone

A mixture of 3-cyanobenzaldehyde (25 g, 0.19 mol), 2-hydroxyacetophenone(25.9 g. 0.19 mol), 38.1 g (0.1 mol) of borax, 250 ml of ethanol, and250 ml of H₂ O was refluxed for four days. When cool the reactionmixture was diluted with H₂ O and extracted with ethyl acetate. Theethyl acetate was dried over anhydrous Na₂ SO₄, filtered, and evaporatedto give a dark, tarry residue. The residue was chromatographed on 1 kgof silica gel, eluting with benzene. From this was isolated 5 g of3'-cyano flavanone as an oil.

NMR Spectrum (CDCl₃): δ 2.95 and 3.0 (s and d, 2H, J=2 Hz) and δ 5.45and 5.60 (d and d, d, J=6 Hz and 10 Hz).

Infrared Spectrum (CHCl₃): 1690 cm⁻¹ (νco), 2240 cm⁻¹ (νcn).

The flavanone was not purified further, but in a similar manner asdescribed in Example 1 was converted to 1.0 g of the hydrochloride saltof 3'-cyano-3-dimethylaminomethyl flavanone as a solid, m.p. 104°-106°C. after recrystallization from 2-propanol-ethyl acetate-petroleumether.

Calcd. for C₁₉ H₁₈ N₂ O₂.HCl: C, 66.56; H, 5.59; N, 8.17. Found: C,65.82; H, 6.08; N, 7.74.

The mother liquors from the recrystallization of the hydrochloride saltwere evaporated and the residue taken up in 500 ml H₂ O and allowed tostand for 1 day. It was extracted twice with ethyl acetate and theextracts dried over anhydrous Na₂ SO₄. It was filtered and evaporated,and the residue purified by preparative tlc to yield 430 mg of thedesired 3'-cyano-3-methylene flavanone as an oil.

NMR Spectrum (CDCl₃): δ 5.2 (1H, m), 6.0 (1H, m), 6.4 (1H, m).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₇ H₁₁ NO₂ : 261.0787. Found:261.0793 (M+).

Infrared Spectrum (CDCl₃): 1680 cm⁻¹ (νco), 2230 (νcn).

EXAMPLE 16 4'-cyano-3-methylene flavanone

In a similar manner as described in Example 1, 4.72 g of 4'-cyanoflavanone, produced as described in Bull. Soc. Chim. France 2248 (1963),was converted to 1.2 g of the hydrochloride salt of4'-cyano-3-dimethylaminomethyl flavanone, m.p. 118°-132° C.

Calcd. for C₁₉ H₁₈ N₂ O₂.HCl: C, 66.56; H, 5.59; N, 8.17. Found: C,65.18; H, 5.94; N, 8.12.

The filtrate from the recrystallization of the hydrochloride salt(2-propanol and ether) was treated in the manner of Example 7 to give200 mg of 4'-cyano-3-methyleneflavanone as an oil.

NMR Spectrum (CDCl₃): δ 5.3 (m, 1H), 6.1 (m, 1H), 6.5 (m, 1H).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₇ H₁₁ NO₂ : 261.0787. Found:261.0768 (M+).

Infrared Spectrum (CHCl₃): 1680 cm⁻¹ (νco), 2230 cm⁻¹ (νcn).

EXAMPLE 17 4',6-dibromo-3-methylene flavanone

A mixture of 25 g (0.135 mol) of 4-bromobenzaldehyde, 29 g (0.135 mol)of 5-bromo-2-hydroxyacetophenone, 51.5 g (0.135 mol) of borax, 180 ml ofethanol, and 300 ml of H₂ O was refluxed for one day. The reactionmixture was cooled, diluted with an equal volume of H₂ O and extractedwith ether. The ether was dried over anhydrous Na₂ SO₄, filtered, andevaporated. This gave 6.67 g of 4',6-dibromo flavanone as a pale yellowsolid, m.p. 167°-168° C.

Calcd. for C₁₅ H₁₀ Br₂ O₂ : C, 47.15; H, 2.64. Found: C, 47.25; H, 2.73.

In a similar manner as described in Example 4, 3.3 g of this flavanonewas converted to 2.6 g of 4',6-dibromo-3-methylene flavanone, isolatedas a pale yellow solid, m.p. 115°-117° C.

Calcd. for C₁₆ H₁₀ Br₂ O₂ : C, 48.76; H, 2.56. Found: C, 49.09; H, 2.67.

NMR Spectrum (CDCl₃) δ 5.2 (m, 1H), 5.9 (m, 1H), 6.4 (m, 1H).

EXAMPLE 18 4'-chloro-7-methoxy-3-methylene flavanone

A mixture of 20 g (0.120 mol) of 2-hydroxy-4-methoxy acetophenone, 16.9g (0.120 mol) of 4-chlorobenzaldehyde, 45.8 g (0.120 mol) of borax, 150ml of ethanol, and 250 ml of H₂ O was refluxed for one day. When cool,the reaction was diluted with an equal volume of H₂ O and extracted withether, then CHCl₃. The organic extracts were combined, dried overanhydrous Na₂ SO₄, and evaporated. The residue was chromatographed on1200 g of silica gel 60, eluting with a gradient of benzene to 19:1 v:vbenzene:MeOH. This gave 3.2 g of 4'-chloro-7-methoxy flavanone as awhite solid, m.p. 121°-122° C.

Calcd. for C₁₆ H₁₃ ClO₃ : C, 66.56; H, 4.54. Found: C, 66.76; H, 4.52.

In a similar manner as described in Example 4, 2.61 g of this flavanonewas converted to 1.16 g of 4'-chloro-7-methoxy-3-methylene flavanone,isolated as a white solid, m.p. 97°-99° C.

NMR Spectrum (CDCl₃): δ 3.8 (s, 3H), 5.2 (m, 1H), 5.9 (m, 1H).

Mass Spectrum (70 eV) m/e: Calcd. for C₁₇ H₁₃ ClO₃ : 300.05524. Found:300.05700 (M+).

Infrared Spectrum (CHCl₃): 1680 cm⁻¹ (νco).

EXAMPLE 19 6-methyl-3-methylene flavanone

In a similar manner as described in Example 4, 4 g of 6-methylflavanone, prepared as described in Justus Liebig's Ann. 587, 1 (1954),was converted to 1.82 g of 6-methyl-3-methylene flavanone, isolated as awhite solid, m.p. 57°-63° C.

Calcd. for C₁₇ H₁₄ O₂ : C, 81.58; H, 5.64. Found: C, 81.25; H, 5.76.

The following compounds comprise preferred embodiments of Formula III:##STR12##

                  TABLE B                                                         ______________________________________                                        3-Methylene Flavanones where R.sup.1, R.sup.4 and R.sup.5 are                 hydrogen and R.sup.3 is phenyl, 2-thienyl or 4-pyridyl                        Example No.  TR Number       R.sup.3                                          ______________________________________                                        20           3742          phenyl                                             21           3928          2-thienyl                                          22           3927          4-pyridyl                                          ______________________________________                                    

EXAMPLE 20 3-methylene-2-phenyl flavanone

In a similar manner as described in Example 4, 4 g of 2-phenylflavanone, prepared as described in Chem. Ber. 94, 241 (1961), wasconverted to 1.30 g of 3-methylene-2-phenyl flavanone, m.p. 165°-167°C., after recrystallization from petroleum ether.

Calcd. for C₂₂ H₁₆ O₂ : C, 84.59; H, 5.16. Found: C, 84.70; H, 5.16.

NMR Spectrum (CDCl): δ 5.0 (m, 1H), 6.6 (m, 1H).

EXAMPLE 21 3-Methylene-2-(2-thienyl) flavanone

A mixture of 56.5 g (0.3 mol) of commercially available2-benzoylthiophene, 24.4 g (0.217 mol) potassium-t-butoxide, 25 mlt-butanol, and 200 ml toluene was heated to reflux, and a solution of 4g (0.0294 mol) 2-hydroxyacetophenone in 75 ml toluene was added dropwisewith stirring. After heating overnight, the mixture was allowed to stir3 days at room temperature; a black tar formed. The black tar waspartitioned between 500 ml 2:1 toluene:ether and 500 ml 1:1 10 percentacetic acid:concentrated hydrochloric acid. The combined organic layerswere washed with brine and dried over sodium sulfate. Filtration andsolvent removal left a black tarry residue which was chromatographed on2 kg of silica gel, eluting with toluene. A 2.4 g portion of a goldenbrown oil, a mixture of the E- and Z-isomers of2'-hydroxy-2-(2-thienyl)chalcone, was obtained. ##STR13##

The chalcone mixture was not further purified but was refluxed overnightin 200 ml of glacial acetic acid containing 50 ml of conc. HCL.Evaporation gave an oil that was chromatographed on 75 g of silica gel,eluting with toluene. A 1.7 g portion of 2-(2-thienyl) flavanone wasisolated as a tan solid, mp 78°-80° C.

Analysis: Calcd. for C₁₉ H₁₄ SO₂ : C, 74.48; H, 4.75. Found: C, 75.39;H, 4.71.

In a similar manner as described in Example 4, 1.5 g of this flavanonewas converted to 1.15 g of 3-methylene-2-(2-thienyl) flavanone, isolatedas a tan solid, mp 164°-166° C.

Analysis: NMR Spectrum (CDCl₃): δ 5.3 (m, 1H), 6.6 (m, 1H).

Mass Spectrum (70 eV) m/e: Infrared Spectrum (CHCl₃): 1680 cm⁻¹ (νco).

Calcd. for C₂₀ H₁₄ O₂ S: 318.0714. Found: 318.0716.

EXAMPLE 22 3-Methylene-2-(4-pyridyl) flavanone

A mixture of 10.1 g (0.055 mol) of commercially available4-benzoylpyridine, 10 ml of t-butanol, 50 ml of dry toluene, and 6.17 g(0.055 mol) of potassium t-butoxide was heated to reflux. A solution of5 g (0.0367 mol) 2-hydroxyacetophenone in 10 ml dry toluene was addeddropwise with stirring, and the mixture refluxed overnight. Aftercooling, the mixture was partitioned between 250 ml of 2:1 toluene:etherand 250 ml of 1:1 (v:v) 10 percent acetic acid:concentrated hydrochloricacid. The aqueous phases were then extracted with chloroform, and thecombined chloroform extracts washed with saturated aqueous sodiumbicarbonate solution, brine, and dried over sodium sulfate. Filtrationand solvent removal left 7.6 g of dark brown oil, which waschromatographed on 250 g of silica gel, eluting with 3:1 v:vchloroform:ethyl acetate. This gave 1.42 g of 2-(4-pyridyl) flavanone asa pale yellow solid, mp 123°-130°.

In a similar manner as described in Example 4, 890 mg of this flavanonewas converted to 525 mg of 3-methylene-2-(4-pyridyl) flavanone, isolatedas a white solid, mp 159°-160° C.

Analysis: NMR Spectrum (CDCl₃): δ 5.0 (m, 1H), 6.4 (m, 1H).

Mass Spectrum (70 eV) m/e: Calcd. for C₂₁ H₁₅ NO₂ : 313.1102. Found:313.1100.

Infrared Spectrum (CHCl₃): 1675 cm⁻¹ (νco).

The following compounds comprise preferred embodiments of Formula IV:##STR14##

                  TABLE C                                                         ______________________________________                                        3-Methylene Chromanones where R.sup.1 and R.sup.2 are hydrogen                and R.sup.3 is naphthyl                                                       Example No.    TR Number     R.sup.3                                          ______________________________________                                        23             3659          1-naphthyl                                       24             3658          2-naphthyl                                       ______________________________________                                    

EXAMPLE 23 3-methylene-2-(1-naphthyl) chromanone

In a similar manner as described in Example 1, 33.4 g of 2-(1-naphthyl)chromanone, as described in J. Indian Chem. Soc. 49, 573 (1972), wasconverted to 9.8 g of the hydrochloride salt of3-dimethylaminomethyl-2-(1-naphthyl) chromanone, isolated as a solid,m.p. 176°-178° C., after crystallization from ethanol-ethyl acetate.

Calcd. for C₂₂ H₂₁ NO₂.HCl: C, 71.82; H, 6.03; N, 3.81. Found: C, 71.96;H, 6.23; N, 3.88.

A 450 ml portion of water was added to 2 g of the compound and stirredfor three days, the compound was then extracted with ethyl acetate. Theethyl acetate extract was dried over anhydrous Na₂ SO₄, filtered andevaporated to give 1.0 g of 3-methylene-2-(1-naphthyl) chromanone as asolid, m.p. 140°-141° C.

NMR Spectrum (CDCl₃): δ 5.0 (m, 1H), 6.4 (m, 1H), 6.6 (m, 1H).

Mass Spectrum (70 eV) m/e: Calcd. for C₂₀ H₁₄ O₂ : 286.0990. Found:286.1021 (M+).

Infrared Spectrum (CHCl₃): 1675 cm⁻¹ (νco).

EXAMPLE 24 3-methylene-2-(2-naphthyl) chromanone

A mixture of 15.6 g (0.1 mol) of 2-naphthaldehyde, 13.6 g (0.1 mol) of2-hydroxyacetophenone, 38.1 g (0.1 mol) of borax, 250 ml of H₂ O, and150 ml of ethanol was refluxed for 4 days. The mixture was cooled,diluted with H₂ O and extracted with ethyl acetate. The ethyl acetatesolution was dried over anhydrous Na₂ SO₄, filtered, and evaporated togive 31 g of an oil consisting chiefly of 2-(2-naphthyl) chromanone. Theoil, not further purified but used directly, was converted in a similarmanner as described in Example 1 into 4.13 g of the hydrochloride saltof 3-dimethylaminomethyl-2-(2-naphthyl) chromanone, isolated as a solid,m.p. 125°-145° C., after recrystallization from acetone.

Calcd. for C₂₂ H₂₀ NO₂.HCl: C, 71.82; H, 6.03; N, 3.81. Found: C, 70.90;H, 6.15; N, 3.61.

The mother liquors from the crystallization of the hydrochloride saltwere evaporated to give a solid residue. The residue was partitionedbetween ethyl acetate and aqueous NaHCO₃ solution. The organic phase wasseparated, dried over anhydrous Na₂ SO₄, filtered and evaporated to give4 g of an oil. It was taken up in 20 ml of methanol and 20 ml of methyliodide and allowed to stand at room temperature for 3 days. The solutionwas then evaporated and the residue partitioned between ether and 10percent aqueous NaOH solution. The ether layer was separated, and theaqueous phase was extracted twice with 100 ml portions of ethyl acetate.The combined organic extracts were washed with H₂ O, 2 percent HCl, H₂O, then dried over anhydrous Na₂ SO₄. Evaporation gave 3.7 g of an oilthat was chromatographed on 400 g of silica gel, eluting with benzene.This gave 1.15 g of 3-methylene-2-(2-naphthyl) chromanone as a solid,m.p. 95°-98° C.

NMR Spectrum (CDCl₃): δ 5.2 (m, 1H), 6.2 (m, 1H), 6.5 (m, 1H).

Mass Spectrum (70 eV) m/e: Calcd. for C₂₀ H₁₄ O₂ : 286.0990. Found:286.0989 (M+).

Infrared Spectrum (CHCl₃): 1675 cm⁻¹ (νco).

Pharmacological Evaluation

The compounds of the present invention were tested for their ability toinhibit the growth of microorganisms by in vitro tests. Generally the invitro tests involve growing representative microorganisms underconditions favorable to such growth and measuring the effectiveness ofone or more compounds in inhibiting or destroying the growth of themicroorganisms. The microorganisms used include representative bacteria,fungi and yeasts.

Compounds which are effective against microorganisms in the abovedescribed in vitro tests may be effective when applied to living tissueor when applied to inanimate objects. The rate of application of acompound effective against a microorganism is dependent upon the potencyand efficacy of the compound. Determination of the rate of applicationfor maximum inhibiting effect is easily achieved by one skilled in theart by evaluating the compound under conditions of use.

The test results summarized in Table D indicate that the compoundsdisclosed and claimed are effective against one or more classes ofmicroorganisms. The compounds are therefore useful in inhibiting thegrowth of microorganisms by applying to a locus inhabited bymicroorganisms an antimicrobially effective amount of such compound.

The test results were obtained as follows. A representative selection ofthe test microorganisms were cultured in a nutrient broth, such as, forexample, Eugon, commercially available from Difco Laboratories, Detroit,Michigan. Bacteria and Candida species were cultured for 24 hours at32°-37° C. and fungi were cultured for approximately one week at 32° C.Inocula were prepared by suspending the growth or spores from a slantculture in sufficient sterile normal (0.9 percent) saline or Eugonbroth, to yield cell or spore concentrations of 1,000,000 cells permilliliter. Anaerobic bacteria were cultured for 48-72 hours at 32°-37°C. in a thioglycollate containing medium, for example, FluidThioglycollate medium or NIH thioglycollate broth, commerciallyavailable from Difco Laboratories.

The compounds of the present invention were tested for activity againstbacteria, yeasts and fungi by the agar incorporation method describedbelow. MA 1347, 3-(dimethylaminomethyl)flavanone (disclosed in U.S. Pat.No. 3,753,985) was used as a comparative standard along with controlsfor each solvent system used.

A series of agar plates containing two-fold dilutions of each compoundto be tested, ranging in concentration from 100 to 3.1 mcg/ml of agarmedium was prepared. Aliquots of each concentration (100, 50, 25, 12.5,6.2, and 3.1) prepared in N,N-dimethylformamide or 50 percent aqueousDMF were added to liquid Eugon agar at 48° C. (one part test solution to100 parts agar). In some cases, tests with further dilutions to 0.125mcg/ml were conducted. The agar medium, containing the 3-methyleneflavanone or chromanone compound to be tested, was then dispensed intopetri dishes and allowed to solidify. The solidified plates were keptovernight at room temperatures to dry the surface of the agar.

The agar plates were then inoculated with a drop of the standardizedcell or spore suspension. Inoculated plates were incubated at 32° C. Theinhibition end-point or minimum inhibitory concentration (MIC) shown inTable D represent the lowest concentration of test compound whichinhibited the growth of the microorganism. Visual determination ofgrowth inhibition was used. Bacteria and yeast end-points weredetermined after 24 hours incubation; molds, dermatophytes and anaerobesafter 48-72 hours incubation. MIC values shown as ranges represent theextremes found in repeat assays. Virtually all such values were writtenplus or minus one dilution, the accepted variation of a two-folddilution assay.

                                      TABLE D                                     __________________________________________________________________________    Minimal Inhibitory Concentration (mcg/ml)                                                         TR 3489                                                                            TR 3667                                                                            TR 3498                                                                            TR 3734                                                                            TR 3673                                                                            TR 3648                                                                            TR 3762                                                                            TR 3666                              MA 1347                                                                             Ex. 1                                                                              Ex. 2                                                                              Ex. 3                                                                              Ex. 4                                                                              Ex. 6                                                                              Ex. 7                                                                              Ex. 8                                                                              Ex.                    __________________________________________________________________________                                                           9                      Staphylococcus aureus                                                                       6.2-12.5                                                                            6.2  ≦3.1                                                                        ≦3.1                                                                        1.56 ≦3.1                                                                        25   12.5 6.2                    ATCC 6538                                                                     S. aureus     6.2- 12.5                                                                           12.5 6.2  3.1  1.56 6.2  25   12.5 6.2                    NRRL B2747                                                                    S. aureus*    25    25   --   --   6.2  --   --   --   --                     NRRL B2747                                                                    Streptococcus faecalis                                                                      50    100  25   25   25-50                                                                              25   100  >100 50                     ATCC 10541                                                                    Sarcina subflava                                                                            12.5  --   --   6.2  3.1  --   --   --   --                     ATCC 7468                                                                     Escherichia coli                                                                            >100  >100 >100 >100 >100 >100 >100 >100 >100                   ATCC 8739                                                                     Proteus vulgaris                                                                            >100  --   --   >100 --   --   --   --   --                     NRRL B123                                                                     Pseudomonas aeruginosa                                                                      >100  >100 >100 >100 > 100                                                                              >100 >100 >100 >100                   ATCC 14502                                                                    Bacteriodides fragilis*                                                                     3.1   1.56 --   --   3.1  --   --   --   --                     ATCC 8482                                                                     Clostridium acetobutylicum*                                                                 ≦100                                                                         --   --   ≦100                                                                        --   --   --   --   --                     ATCC 824                                                                      C. sporogenes*                                                                              6.2   12.5 --   --   ≦0.78                                                                       --   --   --   --                     ATCC 11437                                                                    Propionobacterium acnes*                                                                    12.5  12.5 --   ≦100                                                                        3.1  --   --   >100 --                     ATCC 6919                                                                     Candida albicans                                                                            25-50 50   25   12.5 25   25   >100 100  50                     ATCC 10231                                                                    C. albicans   25    50   25   12.5 25   25   100  100  50                     NRRL Y-477                                                                    C. krusei     12.5-25                                                                             50   25   12.5 25-50                                                                              25   100  50   50                     VM 29B                                                                        C. tropicalis 50-100                                                                              100  50   25   50   25   >100 >100 50                     NRRL Y1410                                                                    C. guillermondii                                                                            50-100                                                                              100  50   25   50   25   >100 100  100                    VM 42                                                                         Trichophyton mentagrophytes                                                                 6.2-12.5                                                                            12.5 6.2  6.2  12.5 6.2  25   12.5 12.5                   ATCC 4807                                                                     T. mentagrophytes                                                                           6.2-12.5                                                                            12.5 6.2  6.2  12.5-25                                                                            6.2  25   12.5 6.2                    TM-2                                                                          T. mentagrophytes                                                                           6.2-12.5                                                                            12.5 12.5 6.2  12.5-25                                                                            12.5 25   12.5 12.5                   CDC                                                                           T. tonsurans  6.2   6.2  --   --   6.2  --   --   --   --                     ATCC 10217                                                                    T. rubrum     ≦3.1-12.5                                                                    --   ≦3.1                                                                        ≦3.1                                                                        12.5 6.2  12.5 25   6.2                    ATCC 10218                                                                    Microsporum gypseum                                                                         6.2-12.5                                                                            6.2-12.5                                                                           6.2  6.2  6.2-12.5                                                                           6.2  25   12.5 6.2                    ATCC 14682                                                                    Aspergillus niger                                                                           50    100->100                                                                           100  25   100  50   >100 100  50                     ATCC 16404                                                                    A. fumigatus  6.2-25                                                                              6.2  --   6.2  50   --   --   50   --                     NRRL 163                                                                      A. fumigatus  50->100                                                                             --   100  50   --   50   >100 --   100                    NRRL 165                                                                      __________________________________________________________________________                        TR 3614                                                                            TR 3654                                                                            TR 3485                                                                            TR 3683                                                                            TR 3649                                                                            TR 3791                                                                            TR 3782                                                                            TR 3783                              MA 1347                                                                             Ex. 10                                                                             Ex. 11                                                                             Ex. 13                                                                             Ex. 14                                                                             Ex. 16                                                                             Ex. 17                                                                             Ex. 18                                                                             Ex.                    __________________________________________________________________________                                                           19                     Staphylococcus aureus                                                                       6.2-12.5                                                                            ≦3.1                                                                        50   6.2  ≦3.1                                                                        12.5 6.2  ≦3.1                                                                        6.2                    ATCC 6538                                                                     S. aureus     6.2-12.5                                                                            6.2  50   6.2  ≦ 3.1-6.2                                                                   12.5 12.5 6.2  6.2                    NRRL B2747                                                                    S. aureus*    25    --   --   --   --   --   --   --   --                     NRRL B2747                                                                    Streptococcus faecalis                                                                      50    50   >100 50   25-50                                                                              100  >100 50   50                     ATCC 10541                                                                    Sarcina subflava                                                                            12.5  12.5 --   12.5 --   --   --   --   --                     ATCC 7468                                                                     Escherichia coli                                                                            >100  >100 >100 >100 >100 >100 >100 >100 >100                   ATCC 8739                                                                     Proteus vulgaris                                                                            >100  >100 --   >100 --   --   --   --   --                     NRRL B123                                                                     Pseudomonas aeruginisa                                                                      >100  >100 >100 >100 >100 >100 >100 >100 >100                   ATCC 14502                                                                    Bacteriodides fragilis*                                                                     3.1   --   --   --   --   --   --   --   --                     ATCC 8482                                                                     Clostridium acetobutylicum*                                                                 ≦100                                                                         ≦100                                                                        --   ≦100                                                                        --   --   --   --   --                     ATCC 824                                                                      C. sporogenes*                                                                              6.2   --   --   --   --   --   --   --   --                     ATCC 11437                                                                    Propionobacterium acnes*                                                                    12.5  ≦100                                                                        --   ≦100                                                                        ≦100                                                                        --   --   ≦100                                                                        ≦100            ATCC 6919                                                                     Candida albicans                                                                            25-50 25   >100 25   50->100                                                                            50   100  50   25                     ATCC 10231                                                                    C. albicans   25    25   >100 25   50->100                                                                            50   100  50   25                     NRRL Y-477                                                                    C. krusei     12.5-25                                                                             25   100  25   50   50   100  25   25                     VM 29B                                                                        C. tropicalis 50-100                                                                              50   >100 50   >100 50   >100 >100 25                     NRRL Y1410                                                                    C. guillermondii                                                                            50-100                                                                              50   >100 50   >100 100  >100 >100 25                     VM 42                                                                         Trichophyton mentagrophytes                                                                 6.2-12.5                                                                            6.2  25   6.2  25   12.5 12.5 12.5 6.2                    ATCC 4807                                                                     T. mentagrophytes                                                                           6.2-12.5                                                                            6.2  25   6.2  25   12.5 25   12.5 6.2                    TM-2                                                                          T. mentagrophytes                                                                           6.2-12.5                                                                            6.2  25   6.2  25   25   50   12.5 6.2                    CDC                                                                           T. tonsurans  6.2   --   --   --   --   --   --   --   --                     ATCC 10217                                                                    T. rubrum     ≦3.1-12.5                                                                    ≦3.1                                                                        12.5 ≦3.1                                                                        12.5-25                                                                            6.2  25   12.5 6.2                    ATCC 10218                                                                    Microsporum gypseum                                                                         6.12-12.5                                                                           6.2  25   6.2  12.5-25                                                                            12.5 50   6.2  6.2                    ATCC 14683                                                                    Aspergillus niger                                                                           50    50   >100 50   100  50   >100 25   25                     ATCC 16404                                                                    A. fumigatus  5.2-25                                                                              12.5 --   12.5 50->100                                                                            --   >100 12.5 25                     NRRL 163                                                                      A. fumigatus  50->100                                                                             100  >100 50   --   100  >100 --   --                     NRRL 165                                                                      __________________________________________________________________________                         TR 3742 TR 3928 TR 3927 TR 3659 TR 3658                                MA 1347                                                                              Ex. 20  Ex. 21  Ex. 22  Ex. 23  Ex.                      __________________________________________________________________________                                                         24                       Staphylococcus aureus                                                                       6.2- 12.5                                                                            0.5     0.5     0.78    ≦3.1                                                                           ≦3.1              ATCC 6538                                                                     S. aureus     6.2-12.5                                                                             0.25-0.5                                                                              0.5     0.78    6.2     6.2                      NRRL B2747                                                                    S. aureus*    25     0.25-1.0                                                                              1.56    1.0     --      --                       NRRL B2747                                                                    Streptococcus faecalis                                                                      50     0.78-1.0                                                                              1.0     3.1     50      50                       ATCC 10541                                                                    Sarcina subflava                                                                            12.5   1.0     1.56    3.1     --      --                       ATCC 7468                                                                     Escherichia coli                                                                            >100   >100    >100    >100    >100    >100                     ATCC 8739                                                                      Proteus vulgaris                                                                           >100   >100    --      --      --      --                       NRRL B123                                                                     Pseudomonas aeruginosa                                                                      >100   >100    >100    >100    >100    >100                     ATCC 14502                                                                    Bacteriodides fragilis*                                                                     3.1    0.125   0.25    0.25    --      --                       ATCC 8482                                                                     Clostridium acetobutylicum*                                                                 <100   1.0     --      --      --      --                       ATCC 824                                                                      Clostridium sporogenes*                                                                     6.2    0.5     0.5     0.5     --      --                       ATCC 11437                                                                    Propionobacterium acnes*                                                                    12.5   0.78-3.1                                                                              1.0     1.0     --      --                       ATCC 6919                                                                     Candida albicans                                                                            25-50  >100    >100    100     >100    >100                     ATCC 10231                                                                    C. albicans   25     >100    >100    100     100     100                      NRRL Y-477                                                                    C. krusei     12.5-25                                                                              >100    >100    100     50      25                       VM 29B                                                                        C. tropicalis 50-100 >100    >100    >100    >100    >100                     NRRL Y1410                                                                    C. guillermondii                                                                            50-100 >100    >100    >100    >100    >100                     VM 42                                                                         Trichophyton mentagrophytes                                                                 6.2-12.5                                                                             >100    12.5    12.5    6.2     12.5                     ATCC 4807                                                                     T. mentagrophytes                                                                           6.2-12.5                                                                             >100    12.5    12.5    12.5    12.5                     TM-2                                                                          T. mentagrophytes                                                                           6.2-12.5                                                                             >100    12.5    25      12.5    25                       CDC                                                                           T. tonsurans  6.2    >100    6.2     6.2     --      --                       ATCC 10217                                                                    T. rubrum     <3.1-12.5                                                                            >100    --      --      6.2     6.2                      ATCC 10218                                                                    Microsporum gypseum                                                                         6.2-12.5                                                                             <3.1->100                                                                             6.2     12.5    6.2     12.5                     ATCC 14683                                                                    Aspergillus niger                                                                           50     >100    >100    >100    >100    >100                     ATCC 16404                                                                    A. fumigatus  6.2-25 >100    100     25      --      --                       NRRL 163                                                                      A. fumigatus  50->100                                                                              >100    --      --      >100    >100                     NRRL 165                                                                      __________________________________________________________________________     *Anaerobic: 48 hours incubation at 35° C. GasPak Anaerobic Jar    

Compounds which compare preferred embodiments of Formula III, Examples20-22, wherein R is phenyl showed increased potency against grampositive microorganisms, e.g., staphylococcus and streptococcusmicroorganisms.

What is claimed is:
 1. A compound of the formula: ##STR15## andpharmacologically acceptable, non-toxic salts thereof wherein: R¹ is amember selected from the group consisting of hydrogen, Br, CH₃ and OCH₃;R² is selected from the group consisting of hydrogen and ##STR16##wherein R⁴ is a member selected from the group consisting of hydrogen,Br, Cl, CH₃, OCH₃, NO₂, N(CH₃)₂, C(CH₃)₃ and CN; R⁵ is selected from thegroup consisting of hydrogen and Cl, with the proviso that when R⁵ isCl, R⁴ is hydrogen or Cl; and R³ is selected from the group consistingof hydrogen, phenyl, 2-thienyl, 4-pyridyl and naphthyl, with the provisothat when R³ is naphthyl, R¹ and R² are hydrogen.
 2. A compound asclaimed in claim 1 wherein R² is ##STR17## and R³ and R⁵ are hydrogen.3. A compound as claimed in claim 2 wherein R¹ is hydrogen.
 4. Acompound as claimed in claim 3 wherein R⁴ is hydrogen.
 5. A compound asclaimed in claim 3 wherein R⁴ is Cl.
 6. A compound as claimed in claim 3wherein R⁴ is Br.
 7. A compound as claimed in claim 3 wherein R⁴ isOCH₃.
 8. A compound as claimed in claim 3 wherein R⁴ is NO₂.
 9. Acompound as claimed in claim 3 wherein R⁴ is N(CH₃)₂.
 10. A compound asclaimed in claim 3 wherein R⁴ is CH₃.
 11. A compound as claimed in claim3 wherein R⁴ is C(CH₃)₃.
 12. A compound as claimed in claim 3 wherein R⁴is CN.
 13. A compound as claimed in claim 1 wherein R¹, R³ and R⁴ arehydrogen; R² is ##STR18## and R⁵ is Cl.
 14. A compound as claimed inclaim 1 wherein R¹ and R³ are hydrogen; R² is ##STR19## and R⁴ and R⁵are Cl.
 15. A compound as claimed in claim 2 wherein R¹ and R⁴ are Br.16. A compound as claimed in claim 2 wherein R¹ is OCH₃ and R⁴ is Cl.17. A compound as claimed in claim 2 wherein R¹ is CH₃ and R⁴ ishydrogen.
 18. A compound as claimed in claim 1 wherein R¹, R⁴ and R⁵ arehydrogen; R² is ##STR20## and R³ is selected from the group consistingof phenyl, 2-thienyl and 4-pyridyl.
 19. A compound as claimed in claim18 wherein R³ is phenyl.
 20. A compound as claimed in claim 18 whereinR³ is 2-thienyl.
 21. A compound as claimed in claim 18 wherein R³ is4-pyridyl.
 22. A compound as claimed in claim 1 wherein R¹ and R² arehydrogen; and R³ is naphthyl.
 23. A method for inhibiting the growth ofmicroorganisms which comprises applying to the locus to be inhibited anantimicrobially effective amount of a compound having the structuralformula: ##STR21## and pharmacologically acceptable, non-toxic saltsthereof wherein: R¹ is a member selected from the group consisting ofhydrogen, Br, CH₃ and OCH₃ ;R² is selected from the group consisting ofhydrogen and and ##STR22## wherein R⁴ is a member selected from thegroup consisting of hydrogen, Br, Cl, CH₃, OCH₃, NO₂, N(CH₃)₂, C(CH₃)₃and CH; R⁵ is selected from the group consisting of hydrogen and Cl,with the proviso that when R⁵ is Cl, R⁴ is hydrogen or Cl; and R³ isselected from the group consisting of hydrogen, phenyl, 2-thienyl,4-pyridyl and naphthyl, with the proviso that when R³ is naphthyl, R¹and R² are hydrogen.
 24. A method as claimed in claim 23 wherein R² is##STR23## and R³ and R⁵ are hydrogen.
 25. A method as claimed in claim23 wherein R¹, R⁴ and R⁵ are hydrogen; R² is ##STR24## and R³ isselected from the group consisting of phenyl, 2-thienyl and 4-pyridyl.26. A method as claimed in claim 23 wherein R¹ and R² are hydrogen; andR³ is naphthyl.